Journal: bioRxiv
Article Title: split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80
doi: 10.1101/2023.03.24.534001
Figure Lengend Snippet: (A) The Gal4 transcription factor binds to UAS sequences to drive transcription, and can be repressed by the binding of Gal80. Gal4 is drawn here as a monomer, but functions as a dimer in vivo. (B) In the original split-Gal4 system, the Gal4DBD and a strong transcriptional activator (VP16 or p65) are each driven by separate enhancers, and reconstituted in cells by leucine zipper domains. Gal80 cannot bind or repress the split-Gal4 complex. (C) In the split-intein Gal4 system, two fragments of the Gal4 protein, each flanked by a split-intein, are independently driven by separate enhancers, and seamlessly trans-spliced to reconstitute a functional, wildtype Gal4 protein, which can be repressed by Gal80.
Article Snippet: The inserts for the original split-Gal4 components (pAW-Zip − -Gal4DBD and pAW-p65-Zip + ) were amplified from pBPZpGAL4DBDUw (Addgene 26233) and pBPp65ADZpUw (Addgene 26234), respectively. pCaSpeR-tub-Gal80 was used to test for Gal80 sensitivity in S2R+ cells.
Techniques: Binding Assay, In Vivo, Functional Assay